Fig. 4: Mapping of folding states within the AesN precursor. | Nature Communications

Fig. 4: Mapping of folding states within the AesN precursor.

From: A cysteine-less and ultra-fast split intein rationally engineered from being aggregation-prone to highly efficient in protein trans-splicing

Fig. 4

a Far UV circular dichroism spectroscopy of the monomeric (blue) and the aggregated (green) species of DTD-(S1A)AesN (8P) in comparison to the fused complex structure MYIDTD-AesN(S1A)-GSH-AesC(N159A)-SVYLN (9P) (gray). b Scheme of the carbene-footprinting reaction to label exposed protein regions using trifluoromethylaryl diazirine (1). c MS-analysis of carbene labeling in AesN tryptic peptides (Pep1–Pep6) in the monomeric form of 8P and the complex structure of MDTD-AesN(S1A)-GSH-AesC(N159A)-SVYLN (10P; 10 µM each) after 2 s of UV irradiation at 77 K using 10 mM aryldiazirine. d SEC-analysis of aggregate and monomer species content of protein constructs (30 µM each) containing partial AesN fragments. For (a), n = 3 technical replicates. Data are presented as mean molar ellipticity corrected for concentration. For (c), n = 5 technical replicates. Data are presented as mean ± s.d. normalized to the total peptide intensity. p-values are derived from a two-way ANOVA Sidak´s multiple comparison test (***p = < 0.0001). Source data are provided as a Source Data file.

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